Cloning and characterization of the autophagy-related gene 6 from the hard tick, Haemaphysalis longicornis

Kawano, Suguru; Umemiya‐Shirafuji, Rika; Boldbaatar, Damdinsuren; Matsuoka, K.; Tanaka, Tetsuya; Fujisaki, Kozo

doi:10.1007/s00436-011-2429-x

Cited by 20 publications

(22 citation statements)

References 29 publications

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“…This process is well conserved in metazoans; however, the study in ticks is still in its infancy. Recently, five ATG genes were characterized in the tick H. longicornis Kawano et al 2011). In this study, we revisited the B. microplus gene index, BmiGI looking for ATG homologues.…”

Section: Discussionmentioning

confidence: 99%

“…However, autophagy in ticks is poorly understood and only few autophagy-related (ATG) genes have been characterized. The HlATG3, HlATG4, HlATG6, HlATG8 and HlATG12 genes were recently reported in the tick Haemaphysalis longicornis (Umemiya et al 2007(Umemiya et al , 2008Umemiya-Shirafuji et al 2010;Kawano et al 2011;Umemiya-Shirafuji et al 2014). Nevertheless, in a study conducted to search for tick homologues of yeast or mammalian ATG genes in eight tick databases, no ATG homologues were found except for H. longicornis and Ixodes scapularis (Malagoli et al 2010).…”

Section: Introductionmentioning

confidence: 84%

“…We conducted a search for ATG homologues in R. microplus, screening the BmiGI database (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus) with the nucleotide sequences of the five ATG genes reported so far from H. longicornis (HlATG3, HlATG4, HlATG6, HlATG8 and HlATG12) plus putative ATG5, ATG7, ATG13 and ATG16 genes from I. scapularis (GenBank accession numbers can be found at Malagoli et al 2010;Kawano et al 2011). Based on sequence similarity we predicted five putative ATG genes (ATG3, ATG4, ATG6 and two ATG8s) (Online Resource 1; Table 1).…”

Section: Atg Homologues In Rhipicephalus Microplusmentioning

confidence: 99%

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Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

et al. 2014

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Rhipicephalus (Boophilus) microplus is an obligate haematophagous arthropod and the major problem for cattle industry due to economic losses it causes. The parasite shows a remarkable adaptability to changing environmental conditions as well as an exceptional ability to survive long-term starvation. This ability has been related to a process of intracellular protein degradation called autophagy. This process in ticks is still poorly understood and only few autophagy-related (ATG) genes have been characterized. The aim of the present study was to examine the ESTs database, BmiGI, of R. microplus searching for ATG homologues. We predicted five putative ATG genes, ATG3, ATG4, ATG6 and two ATG8s. Further characterization led to the identification of RmATG8a and RmATG8b, homologues of GABARAP and MAP1LC3, respectively, and both of them belonging to the ATG8 family. PCR analyses showed that the expression level of RmATG8a and RmATG8b was higher in egg and larval stages when compared to ovary and midgut from adult ticks. This up-regulation coincides with the period in which ticks are in a starvation state, suggesting that autophagy is active in R. microplus.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 84%

Section: Atg Homologues In Rhipicephalus Microplusmentioning

confidence: 99%

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Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

et al. 2014

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“…In the hard tick Haemaphysalis longicornis, endogenous autophagy-related proteins (Atg6 and Atg12) can be detected by western blotting and/or by immunohistochemical analysis of midgut sections. 864,865 It is also possible to detect endogenous Atg3 and Atg8 by western blotting using antibodies produced against the H. longicornis proteins (Umemiya-Shirafuji R, unpublished results). Commercial antibodies against mammalian ATG orthologs (ATG3, ATG5, and BECN1) can also be used for western blotting; however, when the tick samples include blood of a host animal, the animal species immunized with autophagy-related proteins should be checked before use to avoid nonspecific background cross-reactivity.…”

Section: Do Not Distributementioning

confidence: 99%

Guidelines for the use and interpretation of assays for monitoring autophagy

Klionsky¹,

Abdalla²,

Abeliovich³

et al. 2012

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In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

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“…Parthenogenetic H. longicornis ticks (Okayama strain) were maintained by feeding on the ears of Japanese white rabbits (Kyudo, Kumamoto, Japan) for many generations at our laboratory Kawano et al 2011). In this study, the term "unfed" used regarding the state of ticks implies that ticks have not yet attacked new hosts after emerging in the subsequent life stage by hatching or molting.…”

Section: Ticksmentioning

confidence: 99%

RNAi of the translation inhibition gene 4E-BP identified from the hard tick, Haemaphysalis longicornis, affects lipid storage during the off-host starvation period of ticks

et al. 2012

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4E-BP, an eIF4E-binding protein, is well known as a cap-dependent translation inhibitor. Here, the 4E-BP homolog, Hl4E-BP, was isolated and identified from the hard tick Haemaphysalis longicornis. Hl4E-BP transcripts were ubiquitously expressed in the active stages, including the larvae, nymphs, and female adults, and the transcription levels were found to be higher in unfed than engorged ticks. In contrast, the expression levels of non-phosphorylated Hl4E-BP, which is a 13.4-kDa protein detected by the anti-recombinant Hl4E-BP antibody, were the highest in engorged ticks and significantly decreased progressively during the unfed starvation period of ticks. The functional role of Hl4E-BP as a metabolic brake was verified by histochemical observations on the lipid storage in midguts and fat bodies during the starvation period using ticks injected with dsHl4E-BP. The results indicate that Hl4E-BP is highly relevant to the lipid storage of ticks during the non-feeding starvation period. Our results suggest, for the first time, that Hl4E-BP may have a crucial role in the starvation resistance of ticks in an off-host condition via lipid metabolism control, although it was unclear whether Hl4E-BP might be involved in lipid synthesis regulation and/or lipid consumption inhibition.

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Cloning and characterization of the autophagy-related gene 6 from the hard tick, Haemaphysalis longicornis

Cited by 20 publications

References 29 publications

Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Guidelines for the use and interpretation of assays for monitoring autophagy

RNAi of the translation inhibition gene 4E-BP identified from the hard tick, Haemaphysalis longicornis, affects lipid storage during the off-host starvation period of ticks

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